Analisis Sediaan Farmasi

 Analisis Sediaan Farmasi Henry K.S.,M.Si.Apt,

Jadwal Kuliah Minggu

Bahan kajian

I

Validasi metode analisis

II

Validasi metode analisis

III

IV

V

VI

Pemisahan komponen aktif dari bahan pembantu sediaan obat, serta pemilihan pemilihan metode analisis Analisis kuantitatif sediaan obat monokomponen (padat & semi padat) secara spektrofotometri, spektrofluorometri dan elektrokimia Analisis kuantitatif sediaan obat monokomponen (cair dan steril) secara spektrofotometri, spektrofluorometri dan elektrokimia Analisis kuantitatif sediaan obat multikomponen (padat & semi padat) secara spektrofotometri dan spektrofluorometri

VII

Analisis kuantitatif sediaan obat multikomponen (Cair dan Steril) secara spektrofotometri dan spektrofluorometri

VIII

Penentuan dan Identifikasi permasalahan dalam hal analisis

IX

Analisis kuantitatif sediaan obat multikomponen secara KCKT

X

Analisis kuantitatif sediaan obat multikomponen secara KLT KLT dan KG Analisis Sediaan Kosmetika Analisa Sediaan Makanan dan minuman

XI XII XIII XIV

Analisa dalam Sampel Biologis Studi kasus

Daftar Pustaka: • United States Pharmacopoeia • Horwitz, W., and Latimer, G.W., 2005, Official Methods of Analysis, AOAC International, Maryland • Robert V Smith, et. Al, Textbook of Biopharmaceutic  Analysis,1981.  Analysis,1981. • Cahyadi W. , 2006, Analisi 2006, Analisis s dan Aspek Kesehatan Kesehatan Bahan Tambahan Pangan, Pangan, Bumi Aksara: Jakarta • Rohman A., dan I.G. Gandjar, 2007, Metode Kromatografi untuk Analisis Makanan, Makanan , Pustaka Pelajar: Yogyakarta • Mitra S., 2003, Sample Preparation Techniques in  Analytical  Analytical Chemistry , John Willey & Sons: New Jersey

 Analisis Sediaan Farmasi  Analisis = ????? ????? Sediaan Farmasi = ???? C – GMP........ Perlu adanya Validasi

Validation Proses penilaian terhadap parameter  analitik tertentu, berdasarkan percobaan laboratorium, untuk membuktikan bahwa parameter tersebut memenuhi syarat untuk tujuan penggunaannya

The Validation Process

Hardware

Method Validation

Validation Software

System Suitability

Validation (4M)

• Man • Machine • Material • Method

Qualification • Qualification is a subset of the validation process that verifies module and system performance prior to the instrument being placed on-line. • If the instrument is not qualified prior to use and a problem is encountered, the source of the problem will be difficult to identify.

The Validation Timeline

Vendor’s Site

User’s Site

User’s Site

Structural

Qualification

Calibration

and

IQ

and

Software Qualification

OQ

Maintenance,

PQ

System Suitability

Before Purchase

Before Use

After Use

Installation Qualification (IQ) • The IQ process can be divided into two steps: preinstallation and physical installation. • During the preinstallation, all the information pertinent to the proper  installation, operation, and maintenance of  the instrument is reviewed. • Site requirements and the receipt of all of  the parts, pieces, manuals, etc., necessary to perform the installation are confirmed.

Installation Qualification (IQ) • During the physical installation, serial numbers are recorded, and all of the fluidic, electrical, and communication connections are made for components in the system. • Documentation describing how the instrument was installed, who performed the installation, and other miscellaneous details should be archived.

Operational Qualification (OQ) • The OQ process ensures that the specific modules of the system are operating according to the defined specifications for  accuracy, linearity and precision. • This process may be as simple as verifying the module’s self diagnostic routines, or it may be performed in more depth by running specific tests, for  example, to verify detector wavelength accuracy, flow rate, or injector precision.

Performance Qualification (PQ) • The PQ process verifies system performance. • PQ testing is conducted under actual running conditions across the anticipated working range. • In practice, however, OQ and PQ are frequently performed together, particularly for linearity and precision (repeatability) tests, which can be conducted more easily at the system level. • For HPLC, the PQ test should use a method with a well-characterized analyte mixture, column, and mobile phase. A system suitability must be performed. • Proper documentation supporting the PQ process should be archived.

Validation • Process Validation  – Prospective Validation  – Ongoing Validation  – Re-Validation: After change, Periodic,  – Retrospective Validation

• Analytical Method Validation  – Specificity, Linearity, Precision, Accuracy/Recovery, Ruggedness

• Cleaning Validation • Utility System Validation • Computer Validation

What is not  Analytical Method Validation? • Calibration The Process of Performing Tests on Individual System Components to Ensure Proper function

• System Suitability Test to verify the proper functioning of the operating system, i.e., the electronics, the equipment, the specimens and the analytical operations.

HPLC Detector calibration

• Wavelength Accuracy • Linear Range • Noise Level • Drift

Typical System Suitability Test

• Minimum Resolution of 3.0 between the analyte peak and internal standard peaks • Relative Standard Deviation of  replicate standard injections of not more than 2.0%

Method Life Cycle Validation

Development

Optimization

Verification vs. Validation • Compendial vs. Non-compendial Methods  – Compendial methods-Verification  – Non-compendial methods-Validation

Today’s Validation Requirements

ICH/USP

GMPs (legal)

FDA

GMP Validation Parameters •  Accuracy • Specificity • Sensitivity • Reproducibility

FDA Validation Parameters • Accuracy • Precision • Linearity (& Range) • Specificity (& Determination Limit) • Recovery • Ruggedness 1987 FDA Guidelines

ICH/USP Validation Requirements & Parameters ICH

USP

I nter national Conf er ence on H armoni zation 

Specificity

•

Specificity

Linearity and Range

•

Linearity



Accuracy

•

Range



Precision

•

Accuracy

Limit of Detection

•

Precision

 

 

Limit of Quantitation

 – Repeatability

Ruggedness

 – Intermediate Precision

 

Robustness

 – Reproducibility •

Limit of Detection

USP Data Elements Required For Assay Validation

* May be required, depending on the nature of the specific test.

USP Categories • Category 1: Quantitation of major components or  active ingredients • Category 2: Determination of impurities or  degradation products • Category 3: Determination of performance characteristics • Category 4: Identification test

ICH Validation Characteristics vs. Type of   Analytical Procedure

Method Validation for USP • Method validation, according to the United States Pharmacopeia (USP), is performed to ensure that an analytical methodology is accurate, specific, reproducible, and rugged over the specified range that an analyte will be analyzed. • Method validation provides an assurance of  reliability during normal use and is sometime described as the proces of providing documented evidence that the method does what it is intended to do.

KAPAN DILAKUKAN? 1. Pengembangan metode analisis (MA) yang telah ada misalnya untuk: - Matriks sampel yang spesifik - Memperbaiki “Analytical Performance” MA dengan adanya instrument atau teknik baru - MA yang terlalu mahal, memakan banyak waktu & energi - MA alternatif untuk konfirmasi

2. Terhadap MA yang dibuat dari modifikasi metode resmi (standard yang telah dipublikasi secara internasional, regional atau nasional;  jurnal ilmiah yang relevan)

TUJUAN 1. Hasil analisis absah/valid, dapat dipercaya dan dapat dipertanggung  jawabkan secara ilmiah 2. Hasil analisis dapat menunjukkan kesesuaian dengan tujuan pengujian

Accuracy vs precision

Accuracy vs precision What you would like to see!

Accuracy vs precision

• •

Poor accuracy Good precision

Accuracy vs precision

• •

Poor precision Good accuracy

Accuracy vs precision What would you call this?

• • •

Totally hopeless! Poor precision Poor accuracy

So what definitions do these concepts lead us to in the context of assay validation?

 ACCURACY (1) • The accuracy of an analytical procedure expresses the closeness of  agreement between the value which is accepted either as a conventional true value or an accepted reference value and the value found. This is sometimes termed trueness.

 ACCURACY (2) Assay of Drug Substance: a) application of an analytical procedure to an analyte of known purity (e.g. reference material); b) comparison of the results of the proposed analytical procedure with those of a second well-characterized procedure, the accuracy of which is stated and/or defined (independent procedure) c) accuracy may be inferred once precision, linearity and specificity have been established

 ACCURACY (3) Assay of Drug Product: a) application of the analytical procedure to synthetic mixtures of the drug product components to which known quantities of the drug substance to be analysed have been added; b) in cases where it is impossible to obtain samples of  all drug product components, it may be acceptable either to:  – add known quantities of the analyte to the drug product or   – to compare the results obtained from a second, well characterized procedure, the accuracy of which is stated and/or defined (independent procedure)

c) accuracy may be inferred once precision, linearity and specificity have been established.

 ACCURACY (4) Impurities (Quantitation): • Accuracy should be assessed on samples (drug substance/drug product) spiked with known amounts of impurities. • In cases where it is impossible to obtain samples of  certain impurities and/or degradation products, it is considered acceptable to compare results obtained by an independent procedure. • It should be clear how the individual or total impurities are to be determined e.g., weight/weight or area percent, in all cases with respect to the major  analyte.

The Matrix Effect • The matrix effect problem occurs when the unknown sample contains many impurities. • If impurities present in the unknown interact with the analyte to change the instrumental response or  themselves produce an instrumental response, then a calibration curve based on pure analyte samples will give an incorrect determination

 Analytical Method Development • Accuracy: Application of the method to synthetic mixtures of the drug product components to which known quantities of the analyte have been added • Recovery reduced by ~10  – 15%

From: Analytical Method Validation and Instrument Performance Verification, Edited by Chung Chow Chan,Herman Lam, Y.C. Lee and Xue-Ming Zhang, ISBN 0-471-25953-5, Wiley & Sons

Recommended Data • Accuracy should be assessed using a min.

of 9 determinations over a min. of 3 concentration levels covering the specified range (e.g. 3 concentrations/3 replicates each of the total analytical procedure). • Accuracy should be reported as: – % recovery by the assay of known added amount of analyte in the sample or as  – the difference between the mean and the accepted true value together with the confidence intervals

Example: • Taken from: ASEAN Operational Manual for  Implementation of GMP ed. 2000 p.405 • Nine solutions containing different concentrations of ketotifen fumarate reference standard added to ketotifen tablet batch no. 2506VAMG were assayed

Example (continued): Conc. of ketotifen fumarate

Area detected

Recovery (%)

mg/ml

%

0.280

70

1473566

99.32

0.320

80

1677013

99.48

0.360

90

1904848

100.94

0.380

95

1905862

100.51

0.400

100

2091215

100.06

0.420

105

2180374

100.03

0.440

110

2293647

100.07

0.480

120

2518976

101.01

0.520

130

2670144

98.99

Mean (recovery)

: 100.04

Standard deviation

: 0.699

Relative standard deviation (RSD) : 0.699 %

Acceptance Criteria

98.0 – 102.0 % 0.99

Requires a minimum of 5 concentration levels

Method Validation- Linearity

Cara penetapan • Ditetapkan terhadap minimum konsentrasi pada rentang minimum 50 % - 150 % dari kadar analit • Dihitung regresi liniernya dan didapat persamaan regresi: Y = a + bx

RANGE • The specified range is normally derived from linearity studies and depends on the intended application of the procedure. • It is established by confirming that the analytical procedure provides an acceptable degree of linearity, accuracy and precision when applied to samples containing amounts of analyte within or at the extremes of the specified range of the analytical procedure.

Minimum Specified Ranges • for the assay of a drug substance or a finished (drug) product: normally from 80 - 120 % of the test concentration • for content uniformity, covering a minimum of  70 - 130 % of the test concentration • for dissolution testing: +/-20 % over the specified range; e.g., if the specifications for a controlled released product cover a region from 20%, after 1 hour, up to 90%, after 24 hours, the validated range would be 0-110% of  the label claim

Detection limit vs Quantitation limit

‘Know that it’s there’

vs ‘Know how much is there’

Detection limit (means)

Is any of it present?

Is it there?

Quantitation limit  How much of it is present??? 

How much of it is there?

Method Validation- LOD and LOQ Sensitivity • Limit of detection (LOD) – “the lowest content that can be measured with reasonable statistical certainty.”

• Limit of quantitative measurement (LOQ) – “the lowest concentration of an analyte that can be determined with acceptable precision (repeatability) and accuracy under  the stated conditions of the test.”

• How low can you go?

LOD and LOQ Estimated by 1. Based in Visual Evaluations - Used for non-instrumental methods

2. Based on Signal-to Noise-Ratio - 3:1 for Detection Limit - 10:1 for Quantitation Limit

3. Based on Standard Deviation of the Response and the Slope

 Analytical Method Development LOD, LOQ and Signal to Noise Ratio (SNR) LOQ

Signal to Noise = 10:1

Signal to Noise = 3:1

LOD Noise

• Berdasarkan kurva kalibrasi analit Menurut Miller: LOD = 3.SY/X + yb

yb = intersep

LOQ = 10.SY/X +yb S  y / x 

9/23/2013

2 (  ) / N   2  y  y  i ˆ

76

RUGGEDNESS Definisi : Derajat reprodusibilitas hasil uji dari sampel yang sama di bawah kondisi normal, dengan parameter penetapan berbeda, seperti lab, analis, alat, lot pereaksi, hari, waktu & suhu penetapan yang berbeda. Jadi merupakan ukuran reprodusibilitas hasil uji di bawah kondisi normal dari lab ke lab dan dari analis ke analis

Cara penetapan • Sampel dianalisis dari lot sampel homogen, oleh analis berbeda dalam lab berbeda, menggunakan kondisi operasional & lingkungan berbeda tetapi masih dalam spesifikasi yang dipersyaratkan • Ruggedness ditetapkan sebagai fungsi dari variabel penetapan • Ukuran ruggedness MA didapat dari membandingkan reprodusibilitas ini dengan penetapan presisi di bawah kondisi normal

Robustness Small changes do not affect the parameters of the assay

ROBUSTNESS Definisi : Ukuran kemampuan MA untuk tidak terpengaruh oleh perubahan / variasi kecil dari parameter MA yang sengaja dibuat dan memberikan indikasi kehandalan dalam penggunaan normal

Cara penetapan • Dilakukan selama pengembangan MA dan tergantung pada tipe prosedur MA • Bila pengukuran peka terhadap variasi kondisi analitis, maka kondisi analitis tersebut harus dikendalikan • Pada evaluasi robustness, harus ditetapkan parameter kesesuaian sistem (mis: resolusi) untuk menjamin validitas MA tetap terpelihara ketika digunakan